Growing vegetables
M. Zare Mehrjerdi; N. Safari; M. Kharrazi; A. Khadem; A. Sharifi
Abstract
Introduction In addition to providing the necessary energy for photosynthesis, light controls many plant metabolic processes. Nowadays, the use of supplemental light significantly improves the quality of the food product in the conditions of lack of natural light in the autumn and winter seasons. ...
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Introduction In addition to providing the necessary energy for photosynthesis, light controls many plant metabolic processes. Nowadays, the use of supplemental light significantly improves the quality of the food product in the conditions of lack of natural light in the autumn and winter seasons. Light-emitting diodes have been proposed as alternative light sources in controlled agricultural environments. These lamps are the first light sources with the ability to control the light spectrum. Therefore, by controlling the light spectrum and matching the wavelength of LED lamps with the photoreceptors of the plant, the performance and quality of the plant is improved. Cucumber is one of the most important greenhouse plants and its seedlings are generally produced during the autumn and winter seasons, when natural light is relatively low; therefore, the use of supplementary light is required. The use of artificial light sources in fully controlled conditions can change and improve the growth conditions of the plant and also improve its quantitative and qualitative traits. In this research, the aim was to investigate the effect of different qualities of light on the morphophysiological indicators of cucumber seedlings. Materials and Methods This research was performed at the Horticultural Plants Biotechnology Department, Industrial Biotechnology Research Institute of Khorasan Razavi. First, the seeds of the cucumber plant (Cucumis sativus L. var. Officer) were planted in a planting tray, and placed under the light panel with different light treatments. The experimental treatments included four light qualities including white light (6000-6500K) blue light (460-470 nm), red light (625 nm) and combined light (blue + red + white). The amount of photosynthetic photon flux was considered the same in all light treatments, which was equal to 2.75 µol m-2 s-1 CO2 assimilation. Thirty days after planting of seeds, the growth of plants were measured in three stages every 10 days. In each stage, 4 plant samples were selected and then plant height, leaf area, fresh and dry weight of shoots and roots, plant dry matter index and leaf chlorophyll content were measured. Data preparation was done in Excel software, data analysis was done using JMP-8 software and treatment averages were compared using LSD test at 5% probability level. Results and DiscussionThe results of the experiments showed that the quality of light was significantly effective on the growth indicators of Cucumber plants. Plant height in red light and blue light treatments compared to the white light and combined light (red+blue+white with the same intensity) treatments, increased by more than 130% and 60%, respectively. The lowest height was observed in plants grown under combined light, 9 and 14.25 cm after 40 and 50 days of cultivation respectively. The exposure of cucumber seedlings to the red and combined lights recorded the highest and the lowest leaf area in the plants, 618.65 and 377.26 cm2 respectively. Also white light significantly improved the dry weight of the roots. It is worth to mention that the highest plant fresh weight was observed in blue and white lights, 20.9 and 19.5 g respectively. For parameters such as dry weight, dry matter index, and pigment content, the light treatments did not exert a significant effect. However, the utilization of red and blue light, both individually and in combination, positively impacted plant growth. Notably, exposure to red light alone led to a significant increase in leaf surface area, root dry weight, and plant height compared to other light conditions. Previous studies have indicated that red light enhances leaf area, stem length, and fresh weight of plants. Leaves play a critical role in plant photosynthesis and overall growth. Therefore, increasing the leaf area in the plant increases the amount of photosynthesis, growth and development of the plant. In this study, with the increases of the leaf area in the plant and the subsequent increases in the amount of photosynthesis, allocation of dry mater to root increased. The results of the research has been shown that the combination of blue and red wavelengths in pepper, tomato and cucumber seedlings is effective in stimulating plant growth and improving the morphological characteristics under controlled conditions. Blue and red lights can increase the proton flow rate of epidermal cells through the separation mechanism and thus affect leaf development. Blue light directly through the interaction with proton pumps and indirectly through receptors, affects proton pumps by modulating passive ion conduction of potassium and calcium channels. Conclusion According to the results of this research, it was found that exposing the plant to different light quality had different responses in the cucumber plants. Although variables such as dry weight and the amount of photosynthetic pigments were not significantly affected by light quality, however, traits such as plant height, leaf area, root dry weight, and plant fresh weight were affected by light quality. The affected parameters are among the traits that are influenced by the gibberellin hormone and according to the reports related to the effect of light quality on the gibberellin biosynthesis and response to this hormone. It seems that plant action to the quality of light can be attributed to the regulation of this hormone. So it is possible to choose the appropriate light quality in fully controlled conditions according to the production goal and results. In this research, according to the plant leaf rea, root dry weight and plant height, it was determined that white light can be used in the seedling production stage.
Breeding and Biotechnology of Plant and Flower
A. Sharifi; M. Moradiyan; nasim safari; A. Khadem; M. Kharrazi
Abstract
Introduction: Ornamental foliage plants are commonly used for beautifying indoor spaces. Consequently, determining the best method of mass propagation in a short time, is necessary for these plants. For this purpose, an experiment was designed and performed to micropropagate the Singonium ornamental ...
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Introduction: Ornamental foliage plants are commonly used for beautifying indoor spaces. Consequently, determining the best method of mass propagation in a short time, is necessary for these plants. For this purpose, an experiment was designed and performed to micropropagate the Singonium ornamental plant.Materials and Methods: For the first stage, the effect of different plant growth plant growth regulators on the regeneration of nodule explants was evaluated. In this experiment, 1 cm nodal explants were placed in semi-solid MS culture medium containing different concentrations of BA and Kin (0, 1, 2, 3 and 4 mg/l) in combination with 0.2 mg/l IAA. This experiment was performed as a factorial based on a completely randomized design with four replications. The first factor included different types of cytokinin (BA and Kin) and the second factor included different concentrations (0, 1, 2, 3 and 4 mg/l). In the second stage, the effect of different types of culture medium and its different concentrations on plantlet proliferation was investigated. This study was performed as a factorial experiment based on a completely randomized design with five replications. The first factor included the type of culture medium (MS and B5) and the second factor was the different concentrations of culture medium (0.5 and 1). In the third stage of the experiment, the acclimation of in vitro plantlet was investigated. This experiment was performed as a completely randomized design with three replications. The experimental treatments included different culture media: vermiculite, peat moss, vermiculite + peat moss, perlite + peat moss, perlite, coco peat + vermiculite, coco peat + perlite, coco peat and rock wool.Results and Discussion: According to the results of the first experiment, it was found that the use of cytokinin in MS culture medium is effective on plant regeneration. The highest number of regenerated plantlet was observed in culture medium containing 4 mg/l BA with 0.2 mg/l IAA. It is noteworthy that with increasing BA concentration in the culture medium, the plant height decreased, but in contrast, the number of produced plantlets increased. Benzyl aminopurine (BAP) has been introduced as the most important and effective cytokinin in inducing and increasing branching in plants. Research has shown that the use of external benzyl adenine affects plant growth by affecting plant cells or by controlling the accumulation of a number of cytokinin compounds. As a result, the use of cytokinins in culture medium under in vitro culture conditions is necessary to induce and increase cell division. According to the second experimental results, produced plantlets in MS culture medium had higher height, number of roots and root length compared to B5 culture medium. Results also demonstrated that the use of ½ MS in the propagation stage of this plant is appropriate due to the improvement of growth traits such as plantlet height and root length. The use of optimal and suitable culture medium is very effective in the success of plant micropropagation. In the present study, the use of MS culture medium showed better performance compared to B5 culture medium. The appropriate amount of components, better ion strength and more minerals in this culture medium compared to B5 culture medium are probably the factors influencing its superiority in in vitro culture. In the acclimation stage, the use of vermiculite + peat moss culture medium led to 100% adaptation of plants and improvement of growth traits in the plant.Conclusion: The results showed that the type and concentration of cytokinin had a significant effect on the most of the evaluated parameters. With increasing the concentration of cytokinin in the culture medium, the number of regenerated plantlet was increased, but on the other hand, the height of regenerated plantlet also decreased. The use of culture medium containing BA compared to KIN had a more pronounced effect on increasing the number of regenerated plantlets. Application of MS medium compared to B5 medium was more effective in increasing plantlet height, root length, number of roots and number of produced plantlets. Also, half-strength MS medium, increased plantlet height and root length. Therefore, at this stage of propagation, the application of ½ MS culture medium is recommended. In the acclimation stage, it can be stated that the use of vermiculite substrate in combination with peat moss is a suitable option considering 100% compatibility of syngonium plantlets and improvement of growth traits in comparison with other substrates.
Mahdiyeh Kharrazi; A. Sharifi; S. Nejatizadeh; A. Khadem; M. Moradian
Abstract
Introduction: Considering the occurrence of successive droughts in our country and the necessity of optimal water consumption, the use of modern technologies such as closed systems to increase the efficiency of fertilizer and water use will have an effective role in reducing production costs and increasing ...
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Introduction: Considering the occurrence of successive droughts in our country and the necessity of optimal water consumption, the use of modern technologies such as closed systems to increase the efficiency of fertilizer and water use will have an effective role in reducing production costs and increasing yield. The Nutrient Film Technique (NFT) system is one of the hydroponic culture systems in which the nutrient solution is circulated continuously. In this system, a thin layer of nutrient solution will be available to the plant roots, which has a significant effect on reducing fertilizer and water consumption.
Materials and Methods: The present study was conducted to select an optimal system for cultivation of Gerbera plantlets. Plantlets were cultivated in different systems after the acclimation stage: Integrated bedding system as a closed system containing different culture media (Perlite, vermiculite, vermiculite: perlite, cocopeat: perlite and rock wool) and NFT hydroponic system as a closed system. For fertilization, the following fertilizer combination (mM) was used: K2SO4 (0.22), KNO3 (3.62), KH2PO4 (0.71), NH4H2PO4 (0.35), NH4NO3 (0.53), Ca (NO3)2,4H2O (1.48), MgSO4, 7H2O (0.4), MnCl2, 4H2O (0.0049), H3BO3 (0.020), ZnSO4 (0.0061), CuSO4, 5H2O (0.00048), NaMoO4, 2H2O (0.00058), FeSO4, 7H2O (0.0348), Na2EDTA (0.0384). The experiment was conducted in a completely randomized design with six treatments in six replicates. During six months, the growth parameters of the plants including number of leaves, petiole length, fresh and dry shoot weight, leaf chlorosis percentage, leaf necrosis percentage, flower number, flower diameter, flowering stem height, number of days to flowering, end diameter of the stalk and the middle diameter of the stalk were evaluated. Data preparation was done in Excel and data analysis was performed using the Jump 8 software. Mean comparison between treatments was performed with LSD test at 5% probability level and charts were drawn using the Excel program.
Results and Discussion: The results showed that there was a significant difference between treatments for the number of leaves (p <0.01). Seedlings cultivated in the vermiculite culture medium produced the highest number of leaves; however, there was no significant difference between seedlings cultured in vermiculite and vermiculite: perlite media. The lowest number of produced leaves was observed in plantlets grown in the NFT system. Also, the seedlings grown in the rock wool medium had no significant difference with the NFT system. In this regard, the drought stress on plants and the consequent poor growth of plantlets reduced the number of produced leaves. The percentage of necrotic leaves in the NFT system was more than twice the vermiculite culture medium. There was no significant difference between cocopeat: perlite, perlite, rock wool, and NFT systems. Also, the lowest amount of necrotic leaves was found in the vermiculite culture medium, which did not have a significant difference with vermiculite: perlite culture medium. Low levels of leaf necrosis in these treatments indicate the optimal absorption of nutrients and the proper growth of plantlets in these culture media. The results showed that plantlets cultivated in the vermiculite and vermiculite: perlite flowered more quickly than other culture media and also produced more flowers. Plantlets grown in the rock wool medium did not show the desirable performance, which is due to drought stress. Besides, the use of cocopeat in the culture medium did not have a beneficial effect on plants. Measuring the initial electrical conductivity in the cocopeat culture medium also indicates a high EC concentration in this culture medium, which could affect the growth parameters of the plant and lead to poor vegetative growth of plantlets. Since Gerbera plant has bulky roots, the roots of this plant cannot be developed well in the NFT system, so the use of the NFT system is not suitable for Gerbera cultivation. The best performance of plant samples was obtained from vermiculite culture media, but since there were no significant differences between vermiculite and vermiculite: perlite, so to reduce production costs, the use of vermiculite: perlite culture medium in the closed system is recommended for the cultivation of Gerbera plant.
Conclusion: Since Gerbera plant has bulky roots, the roots of this plant cannot be developed well in the NFT system, so the use of the NFT system is not suitable for Gerbera cultivation. The best performance of plant samples was obtained from vermiculite culture media, but since there were no significant differences between vermiculite and vermiculite: perlite, so to reduce production costs, the use of vermiculite: perlite culture medium in the closed system is recommended for the cultivation of Gerbera plant.
A. Sharif; S.M. Kharrazi; S. Nejatizadeh; A. Khadem; M. Moradian
Abstract
Introduction: In traditional agriculture, a high concentration of nutrients is generally available to the plant, and a large amount of it will be leached out and is not available to the plant. But, if properly managed, the nutritional needs of the plant can be identified; it can increase the efficiency ...
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Introduction: In traditional agriculture, a high concentration of nutrients is generally available to the plant, and a large amount of it will be leached out and is not available to the plant. But, if properly managed, the nutritional needs of the plant can be identified; it can increase the efficiency of fertilizer use and also prevent the environmental pollution caused by excessive use of chemical fertilizers in the country. Therefore, the aim of this study was to increase the fertilizer utilization efficiency by choosing optimal fertilizer treatment and its optimum level for acclimation and optimum growth of gerbera tissue culture plantlets.
Material and methods: In the first experiment, different fertilizer treatments with different proportions of NPK were evaluated in a completely randomized design with 10 replications. In the second experiment, the different concentrations of the secondary group treatment (0.5x, 1x and 1.5x) were evaluated to determine the optimum level of fertilizer treatment. This experiment also was conducted in a completely randomized design with 15 replications. At the end of each experiment different parameters such as leaf number, petiole length, shoot dry and fresh weight, main root number, root length, secondary root number, root volume, root dry and fresh weight, chlorophyll a, b and total content were measured. Data preparation was done in Excel and data analysis was performed using the Jump 8 software. Mean comparison between treatments was performed with LSD test at 5% probability level and charts were drawn using Excel program.
Results and discussion: The results showed that the maximum root length, root volume, root dry and fresh weight and secondary root number was obtained from plantlets that was treated by the fertilizer treatment of the second group: K2SO4 (0.22 mM), KNO3 (3.62 mM), KH2PO4 (0.71 mM), NH4H2PO4 (0.35 mM), NH4NO3 (0.53 mM), Ca(NO3)2,4H2O (1.48 mM), MgSO4, 7H2O (0.4 mM), MnCl2, 4H2O (0.0049 mM), H3BO3 (0.020 mM), ZnSO4 (0.0061 mM), CuSO4, 5H2O (0.00048 mM), NaMoO4, 2H2O (0.00058 mM), FeSO4, 7H2O (0.0348 mM), Na2EDTA (0.0384 mM). So among the different fertilizer treatment, application of the fertilizer treatment of the second group was recommended. Calculation of nitrate and ammonium content used in six fertilizer treatments showed that the amount of ammonium used in fertilizer had a significant effect on root number and root dry weight. The maximum root number and root dry weight were observed in the second fertilizer group and then in the first fertilizer group. Urea was not used as a source of nitrogen in the fertilizer formula of these two treatments, while in other fertilizer treatments; urea fertilizer was used in addition to other sources of nitrogen supply. Since urea is converted into ammonium before its absorption by the plant, its application in the fertilizer formula has an effect on the amount of ammonium used in the fertilizer composition. It increases the ratio of ammonium to nitrate in fertilizer composition, which in turn influences root function. The ratio of potassium to phosphorus in the research of Khalaj et al. (4.40) is approximately as the same as the ratio of potassium to phosphorus used in the second group fertilizer treatment (4.50). The results of this ratio were also corresponded to Zheng et al. (4.6), Paradikovic et al (4.4), Hahn et al (4), Savvas and Gizas (4.8).The results of second experiment showed that plantlets grown under 1.5x concentration of second group fertilizer had better growth indices than two other treatments and had higher means in terms of root and shoot dry weight and petiole length. After counting the number of produced leaves at the end of the experiment, signs of leaf necrosis was observed under some treatments. Plantlets treated with 1 and 0.5 x concentrations of the second group treatment showed the highest incidence of leaf necrosis. Application of 1.5 x concentration of the second group treatment resulted in a 40% decrease in leaf necrosis symptoms compared to the other treatments. Also, the highest percentage of healthy leaves was obtained from 1.5 x concentration of the second group treatment, which showed a significant difference with the other treatments.
Conclusion: Generally, for the purpose of acclimation and optimal growth of gerbera tissue culture plantlets, fertilizer treatment of the second group {15(N), 10(P2O5), 30(K2O), 11.10(CaO), 2.10(MgO)} at a concentration of 1.5x is recommended.
S.M. Kharrazi; A. Tehranifar; A. Sharifi
Abstract
Introduction: Success in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Applying different treatments, such as heat treatment and usage of fungicides, can control the microbial contamination and consequently increase the ...
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Introduction: Success in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Applying different treatments, such as heat treatment and usage of fungicides, can control the microbial contamination and consequently increase the percentage of explant survival.
Materials and Methods: This study aimed to investigate the effect of heat treatment and fungicide on reducing the contamination during in vitro culture of narcissus. So, an experiment was done as a factorial experiment in a completely randomized design with two factors, including benomyl concentration in the medium (1 and 2 g/l) and heat treatments (two levels, with and without heat treatment), with 10 replications. In order to sterilizing the plant materials, damaged and infected scales were removed firstly and then bulbs were washed for 30 minutes with running tab water and a few drops of dishwashing liquid. For applying heat treatment, bulbs were divided into two groups. In the first group, heat treatment was not applied and in the second group heat treatment 54 °C was applied for one hour using water bath. After this step, bulbs surface were sterilized by dipping in 70% ethanol for one minute and rinsed with sterile distilled water, followed by immersing in 1.5% sodium hypochlorite solution for 30 min. After sterilization with sodium hypochlorite solution, bulbs were washed three times with sterile distilled water under laminar air flow hood. After sterilization step, bulbs were cut into 32 twin scales explants and cultured in MS medium supplemented with 1 mg/l BA and 0.2 mg/l NAA + benomyl (1 or 2 g/l). After 30 days, the response of explants (number of produced bulbs, percentage of explant survival, percentage of bacterial contamination, percentage of fungal contamination, percentage of browning) was evaluated. Data preparation was done in the Excel program and data analysis was done using JMP-8 software. Mean comparison of the treatments was done by LSD test and finally the charts were drawn using the Excel program.
Results and Discussion: The results showed that increasing the concentration of benomyl in the medium and applying heat treatment had negative effect on regeneration potential of explants, so that the maximum regeneration mean were observed when heat treatment was not applied for explants and medium contains 1 g/l benomyl. Using the heat treatment and application of 2 g/l benomyl in the medium leads to the lowest regeneration amount. On the other hand, evaluating the browning percentage of explants showed that the effect of treatments was significant in this trait. Applying the heat treatment and using 2 g/l benomyl in the medium had severe effect on the increasing of explant browning and the maximum mean was observed in this treatment. But reducing the benomyl concentration in the medium and none application of heat treatment caused the lowest amount of explant browning. Contamination percentage that includes bacterial and fungal contamination is an important parameter in this study. Explants that cultured in the medium containing 1 g/l benomyl and applying heat treatment showed the highest contamination percentage, which contains 21% fungal and 14% bacterial contamination. The lowest percentage of contamination was observed when heat treatment applied and medium contains 2 g/l benomyl. However, this treatment caused the highest percentage of explant browning that lead to reduction of explants regeneration potential. Researches showed that the use of fungicides can help to control tissue culture contamination and according to previous studies, benomyl is the most effective treatment against fungal infection. As benomyl is considered as a systemic fungicide, so it is useful to eliminate the internal fungus. On the other hand, there are some reports about the positive effect of heat treatment on the control of tissue culture contamination. As regards this investigations were done in 1914 to 1997 and then stopped, so it seems that application of this treatment had no sufficient efficiency for contamination control during in vitro condition.
Conclusion: Therefore, by considering the browning, regeneration and contamination percentage, non-application of heat treatment and usage of 1g/l benomyl fungicide in the medium for in vitro culture of Narcissus twin scales explants is recommended.
M. Aghdaei; S.H. Nemati; L. Samiei; A. Sharifi
Abstract
Introduction: Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South ...
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Introduction: Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South American countries, including Bolivia, Colombia, Ecuador and Peru, as well as in countries such as New Zealand and Australia. Pepino is propagated by seed, cutting, and tissue culture methods. Most pepino cultivars are sexually fertile and produce viable seeds, but their seeds have poor germination and high level of heterozygosis causing to highly variable plants. Both mentioned negative aspects have limited the mass production of this plant through seed. In this case, stem cutting is used as the most common way of propagating pepino led to transmission of viral diseases and increasing propagation costs as two main limiting factors of pepino propagation. So, micropropagation systems are a promising tool to produce disease-free clonal plant material with low costs. Therefore, the present study was aimed to assess the effect of different media and plant growth regulators on micropropagation traits of pepino.
Materials and Methods: Three separate experiments were carried out in institute of plant sciences of Ferdowsi University of Mashhad in 2016. Pepino seeds were bought from company of Plant World Seed, UK, were cultivated on MS medium. Grown plants were used as source of providing explants. Four mediums, including MS, ½ MS, SH and B5 were used to determine the best culture medium for shoot regeneration of pepino using single node explant. A factorial experiment was conducted based on a completely randomized design. Some growth properties such as number of shoots, shoot length, number of roots, root length, leaf number and leaf length were evaluated after two and four weeks. In proliferation experiment, MS medium was compared with MS supplemented with different concentrations of BA (0.5, 1 and 2 mg L-1) and Kin (0.5, 1 and 2 mg L-1) applied as combined treatments, and also BA used alone at concentrations of 2, 4 and 6 mg L-1 that was conducted based on a completely randomized design. For rooting of explants, an experiment was conducted based on a completely randomized design containing of two concentrations of IBA (at 0.3 and 0.6 mg L-1) and three concentrations of NAA (at 0.3, 0.6 and 0.9 mg L-1) in MS medium. Some growth properties including root number and length, root density and root quality were evaluated after four weeks
Results and Discussion: Results indicated that micropropagation rate of pepino was affected by culture medium type. The highest shoot length, number of root, root length and leaf number were obtained in MS medium, although statistically there was no significant difference between MS and ½ MS media. The highest number of shoots and leaf length were observed in MS medium, which led to a significant difference with other media (½ MS, SH and B5). Overall, Based on obtained results MS medium was the best culture medium for micropropagation of pepino using single node. In the proliferation experiment, the highest shoot and leaf number and plant color were obtained with using 2 mg L-1 BA + 1 mg L-1 Kin, whereas the highest shoot length and leaf length were observed in the 1 mg L-1 BA + 2 mg L-1 Kin and 1 mg L-1 BA+1 mg L-1 Kin treatments, respectively. Increasing in concentration of BA up to 2 mg L-1 in combination with Kin had a positive effect on shoot proliferation, while applying BA at concentration 2, 4 and 6 mg L-1 alone led to decrease in proliferation. Results obtained from rooting experiment showed that the highest root number, root density and root quality were obtained using IBA at the concentration of 0.6 mg L-1, whereas the highest root length was observed by applying IBA at concentration of 0.3 mg L-1, which led to a significant difference with other treatments. Furthermore, results indicated that the effect of IBA on rooting of pepino microshoots was more than NAA.
Conclusion: Generally, the best results were obtained by MS medium, 2 mg L-1 BA with 1 mg L-1 Kin for shoot proliferation, and IBA at concentration of 0.6 mg L-1 for the rooting of pepino nodal segments.
Ahmad Sharifi; Azadeh Khadem; Maryam Moradian; Mahdiyeh Kharrazi
Abstract
Introduction: Application of stress-tolerant rootstocks is known as one of the effective methods to enhance the productivity of fruit gardens. Pear is one of the four or five major classes of fruits that are produced worldwide. Among pear cultivars, Pyrodwarf rootstock is tolerant to alkaline soil and ...
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Introduction: Application of stress-tolerant rootstocks is known as one of the effective methods to enhance the productivity of fruit gardens. Pear is one of the four or five major classes of fruits that are produced worldwide. Among pear cultivars, Pyrodwarf rootstock is tolerant to alkaline soil and low temperatures of winter. In addition, many cultivars of pear with high productivity could be grafted to this rootstock. These advantages result in widespread culture of pyrodwarf rootstock in pear gardens. The increasing market demand of this rootstock presents an opportunity to investigate alternative methods for efficient production of pyrodwarf. Therefore, this study has been conducted to increase the efficiency of pyrodwarf propagation through tissue culture methods.
Materials and Methods: For this experiment, sterile plantlets of pyrodwarf rootstocks in a same growth phase were used. Plantlets were cultured in different 12 media including MS, WPM, KNOP, modified KNOP, QL and modified QL basal media supplemented with 0.5 mg.L-1 BA and 0.05 mg.L-1 IAA or IBA and solidified with 8 g.L-1 agar. After that, cultures were transferred to the growth room at 25 ± 1°C under a 16/8h light/dark photoperiod (light intensity 30 μmol.m−2.s−1, cool-white fluorescent light). In the next step for rooting optimization, two separate experiments were conducted. In the first, 10 different basal media containing various nutritional compounds were examined for rooting properties of pyrodwarf plantlets. The following experiment was performed to reveal the effects of IAA and active charcoal on the efficiency of plantlets rooting. All the experiments were carried out in acompletelyrandomized design. The study of propagation efficiency of plantlets was arranged in a factorial approach with twofactors including the type of basal media (sixlevels) and auxins (twolevels) with threereplications. The both experiments of plantlet rooting were performed with sevenreplications. Data were analyzed using JMP-8 statistical software and the meanswere compared by using LSD test at 5% probability.
Results and Discussion: All explants were regenerated 15 days after cultivation. The number of buds per explants indicates that nutritional compounds significantly affected bud formation. MS basal media induced the most number of adventitious buds and decreasing amount of nitrogen in the culture medium (either in its ammonium or nitrate form) leads to 50%-decrease in the number of buds induced per explant. Previous reports mentioned that macronutrients have the most significant effect on bud formation of peach hybrid rootstock GF677. In addition, high amount of nitrogen is necessary to achieve the highest height of pyrodwarfin vitro plantlets. In contrast to the basal media, neither bud formation nor height of plantlets did not affected by the type of auxins, IAA and IBA. Also weight of plantlets was similar in all treatments and shows the effect of basal media and auxins were not significant on biomass production. According to the results of rooting experiment, plantlets that were cultured in MS and DKW had higher height than QL and Knop in rooting phase. Also, 1/2 MS-cultured plantlets have the lowest height among all treatments. These results showed that macronutrients have positive effects on the growth of pyrodwarf plantlets as decreasing concentration; the growth of plantlets was inhibited. In contrast to shoot growth, number of roots per explants in 1/2 MS medium was significantly higher than the other media. These results are expected as shoots and roots of plants have different response to environmental stimuli. Data obtained from the following experiment of rooting indicated that IAA improved root induction similar toother functional traits of plants but the presence of both of those compounds in medium have inhibitory effect on the functional traits of plants.
Conclusion: These data suggest that macronutrients and hormonal compounds have impressive impact on the regeneration of pyrodwarf plantlets. In this regard, MS medium supplemented with 0.5 mg.L-1 BA and 0.05 mg.L-1 IAA or IBA induced most number of buds and the regenerated plantlets hadthe highest height in that medium compared to the other treatments. Also, 1/2 MS medium containing 1 mg.L-1 IAA was identified as the best media in case of rooting parameters.
ahmad sharifi; fatemeh keykha; mahboobeh yazdi; abdolreza bagheri
Abstract
Introduction: Lily (Lilium spp.) is a genus of herbaceous flowering plants, which consisted of many beautiful ornamental species with large prominent flowers. Most species are native to the Northern hemisphere temperate, though their range extends into the Northern subtropics. Some specific hybrids of ...
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Introduction: Lily (Lilium spp.) is a genus of herbaceous flowering plants, which consisted of many beautiful ornamental species with large prominent flowers. Most species are native to the Northern hemisphere temperate, though their range extends into the Northern subtropics. Some specific hybrids of Lilium spp. have been developed as cut flower in controlled conditions and in some cases can be grown as pot plant. Propagation rate of lily in natural clonal propagation methods is very low and one year produces of 1-2 bulblets per bulb scale. There is also possibility of disease transmission; so that, tissue culture techniques has provided an efficient method for its micropropagation.
Materials and Methods: In this study, two separate experiments under In vitro conditions the bulblet regeneration from thin cell layer (TCL) explants of Lilium spp. was investigated. In the first experiment, after two months the effect of TCL explants with 1, 3 and 5 mm thickness on MS medium containing 1 mg/l BA, 2ip and kin in combination with 0.5 mg/l NAA on regeneration parameters were assayed. In the second experiment to determine the effect of cultivar and cytokinin types, 3mm thickness TCL explants of five cultivars (Robina, Donato, Nymph, Lessoto and Roxana) were tested on MS medium containing different plant growth regulator (PGR) compounds including BA, kin, 2ip and TDZ at concentration of 1 mg/l in combination with 0.5 mg/l NAA. The regeneration parameters were assayed after four months. In all experiments, the medium was adjusted to pH 5.8 and autoclaved at 121°C for 15 min. All cultures were incubated at 25 ± 2°C with a 16 h photoperiod under cool white flourescent lights (30 µmol/m2).
Results and Discussion: According to the first experiment results, plant growth regulator of BA in all of surveyed parameters except root number was better than other PGRs and explants with 3 mm thickness was the best in all of parameters. The interaction of PGR and explants was significant, however maximum bulblet regeneration was observed in TCL explants with 3 mm thickness in all of PGR treatments (100%). While 1 mm thickness TCL in 2ip and 1 and 5 mm thickness TCL in Kin had the least regeneration percentage. Results revealed that the interaction of explants and medium is a key factor for suitable establishment, regeneration and growth of TCLs. Bulb dormancy is one of the limiting factors in regeneration of bulbous crop species. It seems under In vitro condition explants size and PGR combination of media especially cytokinin affected on breaking of dormancy. Maximum number of leaves and dry weight of bulblets in medium containing BA was significantly higher compared with other treatments. Most of studies confirmed the positive effect of BA on regeneration of lily. The function of cytokinin in plant promoted cell division and differentiation, which lead to growth and maintaining cells in meristematic status.
Result of second experiment showed that cultivar was one of the effective factors on regeneration trait. Oriental lily cultivar "Roxana" had the highest number of roots, bulblets, dry weight and length of plantlets and "Nymph" cultivar showed the lowest percentage of regeneration, dry weight, length of plantlets and rooting obtained. In all of cultivars BA induced more organogenesis percentage and plantlet dry weight, while TDZ induced more rooting percentage.The interaction of cultivar and PGR treatments on percentage of regenerated bulblets and rooting were significant. "Nymph" cultivar had minimum percentage of regeneration and rooting in medium containing TDZ and Kin. Furthermore, "Roxana" cultivar in medium containing BA showed the best dry weight comparison to other treatments.
Conclusion Lily has widely used in the floral industry as a cut flower or potted plant. In recent years, tissue culture was developed as reliable and highly effective method to overcome its limitations of vegetative propagation. The most advantage of this method is high multiplication rate and disease free propagation. In this study, bulblet regeneration of lilium Spp. from TCL explants under in vitro condition was considered as a highly efficient procedure for its micropropagation. With optimization of TCL system some parameters such as exogenously applied plant growth regulators, cultivar, explants types were investigated. Favorable conditions for bulblet regeneration were achieved with 3 mm thickness TCLs in MS medium containing 1 mg/l BA with 0.5 mg/l NAA. This protocol can be used for rapid micropropagation of many cultivars.
ahmad sharifi; Mahdiyeh Kharrazi; Fatemeh Keykha Akhar; Abdolreza Bagheri; Elahe Samari; Maryam Moradiyan
Abstract
Introduction: Gerbera is one of the most important ornamental plants in the world. The importance of Gerbera is due to its beauty, diversity and economically aspects. Traditional propagation methods such as crown division and cutting methods are not suitable for obtaining disease free plants and rapid ...
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Introduction: Gerbera is one of the most important ornamental plants in the world. The importance of Gerbera is due to its beauty, diversity and economically aspects. Traditional propagation methods such as crown division and cutting methods are not suitable for obtaining disease free plants and rapid multiplication. These methods also do not have the capacity to fulfill global demands. Therefore, obtaining efficient protocol for micropropagation of this ornamental plant is necessary.
Materials and Methods: In this study the effect of various factors on in vitro regeneration, proliferation, rooting and acclimation of gerbera capitulum explants were analyzed in four separate experiments. Capitulum explants were first washed with running tap water for 30 min then surface sterilized by dipping in 1.5% sodium hypochlorite solution for 15 min and rinsed with sterile distilled water, followed by immersing in 0.1 % mercuric chloride solution for 10 min. To remove mercuric chloride residue, capitulum was rinsed with sterile distilled water. Subsequent washing was done with sterile distilled water for three times. Sterilization steps were done under laminar air flow hood. For regeneration, eight genotypes of gerbera capitulum explants (Famous, Sunway, Red Pearl, Pink Snow, Popov, Balance, Dune, Eagle)were cultured on solid MS medium containing several cytokynins, BA, TDZ, 2IP or KIN (4 mg/l) in combination with IAA (0.2 mg/l). In proliferation stage, the effect of different concentrations of BA was evaluated on proliferation rate of Sunway regenerated explants. In the rooting stage, Sunway genotype plantlets were cultured on ½ MS medium containing NAA, IBA or IAA (1 mg/l) or ½ MS medium without any hormones. The pH of the medium was adjusted to 5.7-5.8 prior to autoclaving (15 min at 121 oC and 1.5 kg.cm-2 pressure). The cultures were incubated in a growth chamber at 25±2 oC with a 16-h photoperiod (2500-3000 Lux) provided by cool-white fluorescent lamps. For acclimation of rooted plantlets, different substrates used as follow: 1- perlite, 2- perlite: Cocopeat, 3- Cocopeat: peat moss, 4- Cocopeat: peat moss; treated with fungicide.
After 30 days, the response of explants was evaluated for each experiment. Data preparation was done in the Excel program and data analysis was done using JMP-8 software. Mean comparison of the treatments was done by Tukey test and finally the charts were drawn using the Excel program.
Results and Discussion:The results of regeneration stage showed that application of MS media containing kinetin or 2IP did not make an appropriate response to capitulum explants and no regeneration was observed in this condition. The medium containing 4 mg/l BA and 0.2 mg/l IAA indicated the highest percentage of regeneration in all genotypes.
The highest regeneration was observed in Sunway genotype with an average of 21.96%. On the other hand no regeneration was observed in Eagle genotype. In terms of the number of regenerated plantlet, the highest number (61.2) was attributed to the Sunway genotype while no plantlet was recorded for Eagle genotype. No significant differences were also observed between Pink Snow and Dune genotypes.
For the proliferation stage, only Sunway genotype was utilized due to its vigorous growth in comparison to other genotypes. In this stage, the highest (6 regenerated plantlets) and the lowest (1 regenerated plantlet) regeneration rate were observed in MS medium containing 2 mg/l BA and hormone-free medium, respectively. Hormone-free ½ MS medium and ½ MS medium containing 1 mg/l IAA or IBA, indicated the highest rooting rate (100% rooting) while medium containing 1 mg/l NAA showed 55% rooting rate. It seems that the application of NAA in the medium composition had the lowestimpact on the rooting of regenerated plantlets. At the end of the experiment, the highest (90.42%) and the lowest (47.5%) acclimation rate was obtained in peat moss + cocopeat + fungicide medium and perlite medium, respectively.
Conclusions: Generally, for shoot induction of gerbera through capitulum culture, application of MS medium containing 4 mg/l BA and 0.2 mg/l IAA is recommended. It is also concluded that for proliferation stage, the MS medium containing 2 mg/l BA showed the highest rate of regeneration. Using of Hormone-free ½ MS medium is economically affordable. Finally for acclimation of the plantlets, application of peat moss + cocopeat + fungicide medium is recommended.
Ahmad Noroozi; Abdolreza Bagheri; Nasrin Moshtaghi; Ahmad Sharifi
Abstract
Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ...
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Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ofAnthurium genus is a potted perennial plant. Due to having beautiful, attractive and long-life flowers, A. scherzerianum can be used for the production of pot and cut flowers. Tissue culture is suggested as the most commonly method in order to rapid propagation and removing disease in a short period of time. This method also recommended for Anthuriumbecause of problems in classical propagation method of this flower..The three basic propagation methods for Anthuriumare propagation by seed, traditional vegetative and tissue culture.Micropropagation of Anthurium is using forcommercial production.
Materials and Methods: In this study, the effect of plant growth regulators and explants on indirect regeneration of A. scherzerianumdetermined in separate experiments. In the first experiment, callogenesis was done by leaf explants on MS medium containing growth regulators, BA in three concentrations (0.5, 1.25 and 2 mg/l) in combination with
2, 4-D (0.5, 1.25 and 2 mg/l) or NAA (0.5, 1.25 and 2 mg/l) and the combinations of TDZ (0.5, 1.25 and 2 mg/l) with
2, 4-D (0, 0.5 mg/l). In the second experiment, regeneration was done on MS medium containing 0.75 mg/l BA with 0.05 mg/l 2, 4-D and 0.1 mg/l NAA and also in combination with TDZ (0.75mg/l). For rooting, MS medium containing different concentrations of IBA and IAA (0, 0.2 and 1 mg/l) were used. Callus induction, regeneration and rooting experiments were done based on completely randomized design, with 12, 6 and 6 replications, respectively.Data from all the schemes used in this study were analyzed with SAS statistical software. The comparison of means using Duncan's multiple range test was evaluated at the 5% level.
Results and Discussion: Analysis of variance showed that the effect of explant type and hormone combinations was significant on the percentage of callogenesis, callus volume and survival percentage. The interaction effect of explant type and combination of hormones was also significant on percentage of callogenesisand the volume of callus. Means comparisons showed that the highest callogenesis, viability and callus volume were achieved on MS medium containing 2 mg/l of BA and 0.5 mg/l of 2, 4-D. Petiole explants, also produced the highest percentage of callus (95%), survival rate (96%) and callus with dimensions of 6 mm2. Callus formation in leaf vein explants was higher than others. The effect of explant type and hormone combinations on regeneration, number of branches, number of leaves and leaf length was significant.The interaction of explant and hormone combinations on regeneration, number of branches, number of leaves and leaf length was also significant. Moreover, results of regeneration experiment indicated that the maximum number of shoots (6.9) and the maximum shoot length (5 cm), number of leaves (18) and the leaf length (2.8 mm) were achieved in 0.75 mg/l BA mg/l of and 0.05 mg/l 2, 4-D. In this study, petiole explants were also regenerated earlier than leaf explants.The effect of hormone combinations and concentrations was significant on rooting specially on the number of roots and root length.Furthermore, results of rooting experiment revealed that the highest rooting percentage (95%), the maximum number of roots (4.5 per plantlet) and the longest roots (3.5 cm) were produced in the medium containing 0.2 mg/l of IBA. Finally, the rooted plantlets were adapted (90%) in vivo condition by placing them on a mixture of cocopeat and perlite (2:1) substrate.
Conclusion: In this study callugensis, regeneration and rooting of A.scherzerianum’s petiole and leaf explants were studied and different levels of plant growth regulators used for callugensis and regeneration. In this study petiole explants showed the highest callugenesis and regeneration. MS medium containing BA (2 mg/l) and 2, 4-D (0.5 mg/l), was the best for callugenesis. Also the highest percentage of regeneration was observed in medium containing BA (0.75 mg/l) and 2, 4-D (0.05 mg/l). Moreover low concentration (0.2 mg/l) of auxin has a better effect on rooting than high levels (1mg/l) so that the highest rooting percentage was produced in medium containing IBA (0.2 mg/l) and the lowest rooting percentage was produced in medium containing IAA (1 mg/l). Anthurium plantlets acclimized is cocopeat and perlite substrate (2: 1) with 90% acclimation.
Mahdiyeh Kharrazi; Seyyed Hosein Nemati; Ali Tehranifar; Abdolreza Bagheri; Ahmad Sharifi
Abstract
Carnation is considered as the world's third most important cut flower. Tissue culture techniques offer suitable method for micropropagation of this ornamental plant. However, one of the problems during in vitro culture of carnation is vitrification. Ratio of ammonium to nitrate and agar concentrations ...
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Carnation is considered as the world's third most important cut flower. Tissue culture techniques offer suitable method for micropropagation of this ornamental plant. However, one of the problems during in vitro culture of carnation is vitrification. Ratio of ammonium to nitrate and agar concentrations in the medium affect this phenomenon. Therefore, in this study the effect of these factors on the rate of proliferation and the vitrification of carnation (Dianthus caryophyllus L.) cultivar Innove Orange Bogr, was evaluated. In this investigation lateral buds were cultured on MS medium containing 1 mg/l BA, 0.1 mg/l NAA and with different concentrations of agar and different ratio of ammonium to nitrate. The results showed that increasing in the agar concentration to 12 g/l lead to decreasing the rate of vitrification but regeneration also declined. Increase of agar concentration cause limitation in nutrient absorption by plants. Also, decrease in the ratio of ammonium to nitrate in the medium reduces the amount of vitrification, but did not result in adverse effects on plant regeneration rates. Multiple regressions showed that the effect of ammonium to nitrate ratio on vitrification was higher than agar concentration. So by considering the amount of shoot regeneration and vitrification, to obtain the most normal shoots, the concentration of 10 g/l agar with ammonium to nitrate ratio 1:6, is recommended.
Mahdiyeh Kharrazi; Seyyed Hosein Nemati; Ali Tehranifar; Ahmad Sharifi; Abdolreza Bagheri
Abstract
Abstract
Carnation (Dianthus caryophyllus L.) is the third most important cut flowers in the world. Tissue culture techniques offer an efficient method for micropropagation of this ornamental plant. In present work effects of Kinetin (Kin) and Benzyl Adenine (BA), on shoot multiplication and hyperhydricity ...
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Abstract
Carnation (Dianthus caryophyllus L.) is the third most important cut flowers in the world. Tissue culture techniques offer an efficient method for micropropagation of this ornamental plant. In present work effects of Kinetin (Kin) and Benzyl Adenine (BA), on shoot multiplication and hyperhydricity of four carnation cultivars (Prado Aquila Kgr, Skimo Mogr, Mondeo Kgr and Innove Orange Bogr) were studied. Explants from nodal segments were cultured on MS medium supplemented with different concentrations of BA (1, 2, 3 and 4 mg/l) and Kin (1, 2, 3 and 4 mg/l) in combination with 0.2 mg/l NAA, 30 g/l sucrose and 8 g/l agar. Rooting of regenerated shoots was done in the MS medium supplemented with 1 mg/l NAA. Results indicated that there is a significant difference among cultivars shoot regeneration, Eskimo and Prado Aquila Kgr with 3.2 and 1.5 shoots show the highest and lowest regeneration rate, respectively. Increasing the concentration of cytokinin from 1 mg/l to 4 mg/l lead to increase regenerated shoot number from 1.7 to 2.4 shoots per explant and increase hyperhydricity from 12% to 54%. In addition high concentration of cytokinin, especially BA, decreased height of regenerated shoots. Based on hyperhydricity percentage of regenerated shoots, there was a significant difference between cultivars and cytokinins. Mondeo Kgr and Prado Aquila Kgr showed highest (44%) and lowest (23%) hyperhydricity, respectively and explants cultured in medium supplemented with BA caused more (40%) hyperhydricity than Kin (26%). Results of present work showed that by increasing the concentration of cytokinin specially BA, multiplication will be increased but also it will increase the hyperhydricity of plantlets and consequently it will lead to death of them. By considering the amount of multiplication and frequency of hyperhydricity for obtaining the highest number of normal shoots, using of MS medium containing 1 mg/l BA in combination with 0.2 mg/l NAA is suggested.
Keywords: Carnation, Benzyl Adenine, Kinetin, Hyperhydricity, Shoot multiplication, Micropropagation
Amirghaffar Ghaffar Shahriari; Abdolreza Bagheri; Ahmad Sharifi; Nasrin Moshtaghi
Abstract
Abstract
Alstroemeria is one of the most important cut flowers in the world which is multiplicated by rhizome explants in in vitro conditions . The major problem in using rhizome explants is fungal and bacterial contamination which inhibit the preparation of strill explants. Therefor in this experiment, ...
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Abstract
Alstroemeria is one of the most important cut flowers in the world which is multiplicated by rhizome explants in in vitro conditions . The major problem in using rhizome explants is fungal and bacterial contamination which inhibit the preparation of strill explants. Therefor in this experiment, the effect of disinfecting compounds like sodium hypochlorite, mercury chloride, nano silver particles, the antibiotics of streptomycin, penicillin, cefotaxime and carbendazim fungicide were examined on controlling of contamination. After disinfection, the explants were cultured on MS medium supplemented with 2 mg/L BA and 0.2 mg/L NAA. Contamination percentage was measured after three weeks. The results showed that the treatment of plants with 0.4% carbendazim fungicide and disinfection by 70% ethanol for one minute and %3 sodium hypochlorite for 20 minutes for caralis cultivar, and disinfection by 70% ethanol for one minute and %3 sodium hypochlorite for 20 minutes follow by culturing in medium with 200 mg/l either of streptomycin and penicillin in Bordeaux cultivar had the lowest contamination percentage. Contamination was not controlled by nano silver particles in studied plants.
Keywords: Alstroemeria, sodium hypoclorite, mercury chloride, antibiotics, carbendazim
